2.4. Magnetic-Activated Cell Sorting (MACS)

The magnetic sorting of HT1080-Qt cells after 24 h of incubation with 1 mM FAS (Sigma-Aldrich) was performed using a magnetic separation kit (Miltenyi Biotec, Bergisch Gladbach, North Rhine-Westphalia, Germany) [41]. The cells were thoroughly washed with DPBS from FAS, detached from the plastic by trypsinization, centrifuged (500× g, 5 min), and resuspended in 1.5 mL of DPBS containing 2% FBS. The number of cells was counted using an automatic cell counter. MS column was placed into OctoMACS Separator and equilibrated with 0.5 mL PBS. The cell suspension was applied to a magnetic column, and free-passing cells were collected in a 15 mL tube. The column was washed twice with 1 mL of DPBS containing 2% FBS, removed from the separator’s magnetic field, and transferred in a new 15 mL tube. Next, 1 mL of DPBS with 2% FBS was added to the column, and the cells retained in the column were eluted using a plunger. Then, the cells were centrifuged (500× g, 5 min), resuspended in a complete DMEM, and counted. Control HT1080 cells were processed according to the same protocol.